Pectin from apple

C. cellulovorans 743B (ATCC35296) was grown anaerobically as previously described (Sleat et al. 1984 (link)), differing only in the carbon source, which was replaced by 0.3% (w/v) glucose (Nacalai Tesque, Kyoto, Japan), 0.3% (w/v) xylan from beechwood (Sigma, MO, USA), 0.3% (w/v) pectin from apple (Sigma), or 0.3% (w/v) LBG from Caretonia silliqua seeds (Sigma).

β-cyclodextrin (>98%), pectin from apple (M_V: 9000; 73.9% esterification), sulfuric acid (96.5%), poly(vinyl alcohol) (M_wca. 13 000; 98.0–98.8 mol% hydrolysis), dimethylformamide (DMF) (99%), acetone (99.7%) and dialysis tubing cellulose membrane (molecular weight cut-off: 14 000 Da MWCO) were obtained from Sigma-Aldrich (Germany). Chitosan with acetylation degree of 15 mol% (M_wA: 87 × 10³ g mol⁻¹) was purchased from Golden-Shell Biochemical (China). Benzyl alcohol (99%) was obtained from Merck KGaA (Germany).
Benzene (ben) (>99.7%) and xylene (xyl) (mixture of isomers >97%) were purchased from Merck KGaA (Germany) and toluene (tol) (>99.8%) was obtained from Lab-Scan (Poland). Pyrene (pyr) (GC grade > 97%), benzo(b)fluoranthene (B(b)F) (HPLC grade > 98%) and benzo(a)pyrene (B(a)P) (>96%) were purchased from Sigma-Aldrich (Germany).
Acetonitrile and methanol, HPLC grade, were purchased from Fisher Scientific (UK) and Sigma-Aldrich (Germany), respectively. Deionized water was obtained using a Millipore© system. All reagents were used without further purification.

Agarose (A6013), L-Alanine (A7627), Benzoic acid (242381), 3,3′-diaminobenzidine (D5905), Folic acid (F7876), Formic acid (F0507), D-(–)-Fructose (F0127), D-(+)-Glucose (BDH0230), Glycine (G7126), Horseradish peroxidase (P6782), DL-Malic acid (M0875), D-Mannitol (M4125), N-Z-Soy® Peptone (P1265), Pectin from apple (P8471), Phosphoric acid, D-(–)-Quinic acid (138622), Starch from rice (S7260), and Sucrose (50389) were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO). Acetonitrile (AX0145) was obtained from EMD Millipore (Billercia, MA), Citric acid anhydrous (A940) from Fisher Scientific (Fair Lawn, NJ), Sorbitol (V045-07) from J. T. Baker – Mallinckrodt Baker, Inc. (Phillipsburg, NJ) and Proflo Premium Quality Cottonseed-derived Protein Nutrient (069061) from Trades Protein, Southern Cotton Oil Company (Memphis, TN). All solvents were of HPLC grade and water was of Milli-Q quality (Millipore Corp., Bedford, MA).

Pectin from apple (76282, degree of esterification—DE: 70–75%), citrus peel (P9135) and methyltrimethoxysilane (MTMS, 98%) were supplied from Sigma-Aldrich (Steinheim, Germany). Hexamethyldisilazane (HMDZ, 98.5%) from abcr GmbH (Karlsruhe, Germany), and ammonium hydroxide (NH₄OH, puriss. p.a.) from PanReac AppliChem (ITW Reagents, Barcelona, Spain). Tetraethyl orthosilicate (TEOS, 98%) was bought from Acros Organics and Ethyl acetate (EtOAc ≥ 99.8%) and acetic acid (CH₃COOH, glacial, analytical reagent grade) were supplied from Fisher Chemical and Fisher Scientific, all belonging to the Thermo Fisher Scientific group (Waltham, MA, USA). Ethanol absolute was purchased from VWR (Radnor, PA, USA). The water used was purified through a Millipore system. CO₂ (purity > 99%) was supplied by Nippon Gases (Madrid, Spain). Commercial hydrophilic cotton fibers were used as the reinforcement matrix, previously aligned in the lengthwise direction. All the remaining chemicals used were of analytical grade.

Polygalacturonic acid, pectin from citrus peel, and pectin from apple were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sodium polygalacturonate was purchased from Shanghai Yuanye Bio-Technology Co., Ltd (Shanghai, China). D-Galacturonic acid sodium salt (purity, about 95%) was obtained from Sigma-Aldrich. All other chemicals and reagents were of analytical grade. The genome of Echinicola rosea JL3085^T harbored multi-gene polysaccharide utilization loci (PUL) systems involved in the degradation of pectin. This study used Escherichia coli DH5α and E. coli BL21 (DE3) for plasmid construction and as the hosts for gene expression, respectively.